血管性痴呆大鼠模型学习记忆障碍与海马半胱氨酸蛋白酶1蛋白的表达:空白对照及量化盲法评估

Learning and memory disorder and the expression of cysteine protease 1 in hippoc ampus of rats with vascular dementia:blank-control and quantified blind evaluation

目的:定量测定血管性痴呆大鼠的学习记忆成绩,同时观察海马半胱氨酸蛋白酶1蛋白在血管性痴呆大鼠海马中的表达及其意义。方法:实验于2004-01/11在中国医科大学动物实验室进行,取健康雄性SD大鼠100只,随机分正常对照组(n=28,仅分离双侧颈总动脉)、假手术组(n=28,仅麻醉,不做手术),和血管性痴呆组(n=44)。采用血管阻断的方法,复制大鼠血管性痴呆模型,每组随机取10只大鼠,采用水迷宫实验进行行为学检测,定量测定其学习记忆成绩。每组剩余大鼠在缺血后24,48,72,96,120h及7d处死,用免疫组化方法检测海马区半胱氨酸蛋白酶1蛋白表达水平的变化。结果:100只大鼠死亡4只,进入结果分析96只。①水迷宫学习成绩:血管性痴呆组大鼠游完全程的时间、进入盲端的次数及时间[(3.78±1.65)min,(5.21±1.56)次,(1.90±1.38)min]明显多于正常对照组及假手术组[(0.68±0.12)min,(1.13±0.21)次,(0.21±0.08)min;(0.76±0.13)min,(0.98±0.32)次,(0.31±0.09)min]。②水迷宫记忆成绩:血管性痴呆组大鼠游完全程的时间、进入盲端的次数及时间[(2.98±1.05)min,(5.62±2.11)次,(1.78±0.84)min]明显多于正常对照组及假手术组[(0.24±0.16)min,(0.48±0.73)次,(0.04±0.02)min;(0.35±0.20)min,(0.52±0.80)次,(0.10±0.02)min]。

AIM:To estimate the learning and memory results in rats with vascular dementia quantitatively,and observe the expression of cysteine protease 1 in hippocampus and the significance. METHODS:The experiment was performed in the Animal Laboratory of Capital Unive rsity of Medical Sciences from January to November 2004.A total of 100 SD rats w ere divided into control group(n=28,only separated bilateral common carotid arte ry),sham operation group(n=28,only received anesthesia not operation) and vascul ar dementia group(n=44).The method of vascular blocking was used to establish th e vascular dementia models.Ten rats were selected from each group to detect the praxiology by suing Water Maze test,and the learning and memory results were est imated quantitatively.The rest rats in each group were killed at hours 24,48,72, 96,120 and day 7 following ischemia.The expression of cysteine protease 1 in hip pocampus was detected by using immunohistochemical method. RESULTS:Four of 100 rats died and a total of 96 were involved in the result.① Learning result indicated:the swimming time and number of error in vascular deme ntia group[(3.78±1.65) minutes,(5.21±1.56) times,(1.90±1.38) minutes] increas ed obviously as compared with that of control group[(0.68±0.12) minutes,(1.13± 0.21) times,(0.21±0.08) minutes] and sham operation group[(0.76±0.13) minutes, (0.98±0.32) times,(0.31±0.09) minutes].②Memory result indicated:the swimming time and number of error in vascular dementia group[(2.98±1.05) minutes,(5.62± 2.11)times,(1.78±0.84) minutes] increased obviously as compared with that in co ntrol group[(0.24±0.16) min utes,(0.48±0.73) times,(0.04±0.02) minutes] and s ham operation group[(0.35±0.20) minutes,(0.52±0.80) times,(0.10±0.02) minutes ].③After cerebral ischemia for 24 hours,the expression of cysteine protease 1 i n vascular dementia group was 82±12,and reached to the peak at the third day,wh ich was significantly higher as compared with that in control group(17±3) and s ham operation group(20±3). CONCLUSION:Obvious learning and memory disorder occurs in rats with vascular d ementia.The expression of cysteine protease 1 expresses significantly in hippoca mpus.As a gene for inhibiting the apoptosis,cysteine protease 1 can protect the neurons in hippocampus,which has close relation with learning and memory of rats with vascular dementia.